Ifn γ elisa

Manufactured by BioLegend

Sourced in United States

The IFN-γ ELISA is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) used to measure the concentration of interferon-gamma (IFN-γ) in biological samples. This assay utilizes antibodies specific for IFN-γ to detect its presence in the sample.

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Lab products found in correlation

Duoset elisa kit, by R&D Systems (1 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions) Non essential amino acids (neaa), by Merck Group (1 mentions) Xtt assay, by Roche (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Polyacrylamide gel, by Bio-Rad (1 mentions) Recombinant human il 15, by BioLegend (1 mentions) Dbco nhs ester, by Vector Laboratories (1 mentions) Sulfo cyanine7 nhs ester, by Lumiprobe (1 mentions) Recombinant mouse scil 12, by Miltenyi Biotec (1 mentions) Recombinant human il 2, by Thermo Fisher Scientific (1 mentions) Human t helper cytokine panel legendplex bead based immunoassay, by BioLegend (1 mentions) Tnfα elisa, by BioLegend (1 mentions) Il17a elisa, by BioLegend (1 mentions) Il 13 elisa, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Mouse IFN-γ ELISA kit Human and mouse IFN-γ ELISA kits LEGEND MAX™ Mouse IFN-γ ELISA Kit ELISA MAX Deluxe Set Human IFN-γ Human ELISA kits for IFN-γ ELISA kits for IFN-γ ELISA MAX standard set mouse IFN-γ IFN-γ ELISA kit LEGEND MAX™ Human IFN-γ ELISA Kit Mouse IFN-γ ELISA MAX

14 protocols using ifn γ elisa

NK Cell-Mediated Glioblastoma Differentiation

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To differentiate with serum, the brain CSCs were cultured in DMEM medium supplemented with 10% FBS for a period of 4 weeks. To differentiate with NK cell supernatants, NK cells were left untreated or treated with the combination of anti-CD16mAb (3ug/ml) and IL-2 (1000 units/ml) for 18- 24 hours before the supernatants were removed and used in differentiation experiments. The amounts of IFN-γ produced by activated NK cells were assessed with IFN-γ ELISA (Biolegend, CA). Serum-differentiated XO2GB (XO2GB-S) were further selected for differentiation with the NK cell supernatants. Differentiation of XO2GB-S with NK cell supernatants was conducted with gradual daily addition of increasing amounts of NK cell supernatants. XO2GB-S required on average a total of 0.035pg of IFN-γ containing supernatants from IL-2+anti-CD16mAb treated NK cells per tumor cell during a 7 day treatment, whereas XO2GB CSCs required 0.070pg of IFN-γ containing supernatants from IL-2+anti-CD16mAbmAb treated NK cells per tumor cell for 7-10 days to promote differentiation and resistance to NK cell mediated cytotoxicity. Initially 1X10⁶ tumor cells were cultured and treated with NK supernatants for differentiation. Afterwards, target cells were rinsed with 1X PBS, detached and used for experiments.

Tseng H.C., Inagaki A., Bui V.T., Cacalano N., Kasahara N., Man Y.G, & Jewett A. (2015). Differential Targeting of Stem Cells and Differentiated Glioblastomas by NK Cells. Journal of Cancer, 6(9), 866-876.

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Cytokine Quantification in Cell Cultures

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Supernatants from PBMCs and sorted cell co-cultures were used to determine levels of IFNγ ELISA (Biolegend), IL-2HS (sensitivity 0.4 pg/ml) and IFNγHS (sensitivity 0.99 pg/ml) (both from eBioscience). Plasma IL-27 levels were measured using Duoset ELISA kit (R&D Systems).

Garg A., Trout R, & Spector S.A. (2017). Human Immunodeficiency Virus Type-1 Myeloid Derived Suppressor Cells Inhibit Cytomegalovirus Inflammation through Interleukin-27 and B7-H4. Scientific Reports, 7, 44485.

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Enhancing NK Cell Cytotoxicity Against OSCSCs

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NK cells were purified from healthy donor’s PBMCs as described above. NK cells were left untreated or treated with IL-2 (1000 units/ml) and anti-CD16 mAb (3 μg/ml) with or without monensin (1:1300) for 24 h as per manufacturer’s recommendation. Afterward, the supernatants were harvested from NK cells and the amounts of IFN-γ produced were measured with IFN-γ ELISA (BioLegend, CA, USA). NK cells were then fixed with freshly prepared 2% paraformaldehyde for 15 min and then washed three times with 1× PBS. Thereafter, fixed NK cells were added to OSCSCs at an effector to target ratio of 0.75–1. After 5 days of treatment, NK cells were removed from culture and OSCSCs were used for experiments.

Tseng H.C., Bui V., Man Y.G., Cacalano N, & Jewett A. (2014). Induction of Split Anergy Conditions Natural Killer Cells to Promote Differentiation of Stem Cells through Cell–Cell Contact and Secreted Factors. Frontiers in Immunology, 5, 269.

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NK Cell Supernatant Induces Stem Cell Differentiation

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Human NK cells were purified from healthy donor’s PBMCs as described above. NK cells were left untreated or treated with anti-CD16 mAb (3 μg/ml), IL-2 (1000 units/ml), or a combination of IL-2 (1000 units/ml) and anti-CD16 mAb (3 μg/ml) for 18–24 h before the supernatants were removed and used in differentiation experiments. The amounts of IFN-γ produced by activated NK cells were assessed with IFN-γ ELISA (BioLegend, CA, USA). Differentiation of OSCSCs was conducted with gradual daily addition of increasing amounts of NK cell supernatant. On average, a total of 1500 pg of IFN-γ containing supernatants obtained from IL-2 + anti-CD16 mAb-treated NK cells was added for 5 days to induce differentiation and resistance of OSCSCs to NK cell-mediated cytotoxicity. DPSCs and SCAP cells required on average a total of 3600 pg of IFN-γ containing supernatants obtained from IL-2 + anti-CD16 mAb-treated NK cells during a 7-day treatment, whereas MP2 tumors required a total of 7000 pg of IFN-γ containing supernatants from IL-2 + anti-CD16 mAb-treated NK cells for 7 days to promote differentiation and resistance to NK cell-mediated cytotoxicity. Afterward, target cells were rinsed with 1× PBS, detached, and used for experiments.

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Differentiation of Oral Cancer Stem Cells

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Human NK cells were purified from healthy donor's PBMCs as described above. NK cells were left untreated or treated with anti-CD16mAb (3 ug/ml), IL-2 (1000 units/ml) or a combination or IL-2 (1000 units/ml) and anti-CD16mAb (3 ug/ml) for 18–24 hours before the supernatants were removed and used in differentiation experiments. The amounts of IFN-γ produced by the activated NK cells were assessed with IFN-γ ELISA (Biolegend, CA). Differentiation of OSCSCs was conducted with gradual daily addition of increasing amounts of NK cell supernatant. On average a total of 1000 pg of IFN-γ containing supernatants obtained from IL-2+anti-CD16mAb treated NK cells was added for 4 days to induce differentiation and resistance of OSCSCs to NK cell mediated cytotoxicity. DPSCs required on average a total of 3600 pg of IFN-γ containing supernatants obtained from IL-2+anti-CD16mAb treated NK cells during a 4 day treatment to promote differentiation and resistance to NK cell mediated cytotoxicity. Afterwards, target cells were rinsed with 1X PBS, detached and used for experiments.

Tseng H.C., Cacalano N, & Jewett A. (2015). Split anergized natural killer cells halt inflammation by inducing stem cell differentiation, resistance to NK cell cytotoxicity and prevention of cytokine and chemokine secretion. Oncotarget, 6(11), 8947-8959.

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NK Cell-Induced Tumor Cell Differentiation

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Human NK cells were purified from healthy donors' PBMCs as described above. NK cells were left untreated or treated with a combination of IL-2 (1000 units/mL) and anti-CD16mAb (3 μg/mL), (a condition to induce split anergy in NK cells) for 18- 24 hours before the supernatants were removed and used in differentiation experiments. The amounts of IFN-γ produced by activated split-anergized NK cells were assessed with IFN-γ ELISA (Biolegend, San Diego, CA, USA). Differentiation of OSCSCs and MP2 cells was conducted with an average total of 1500-2000 pg and 4000-7000 pg of IFN-γ from IFN-γ containing supernatants, respectively, over 4-5 days as described previously 18 (link). Differentiation of A549 and A375 cells was conducted with an average total of 6000 pg and 15000 pg of IFN-γ containing supernatants, respectively over the course of 5 days. Differentiation of X02GB required 35000 pg of IFN-γ containing supernatants as described previously 57 . Initially, 1X10⁶ tumor cells were cultured and treated with split-anergized NK supernatants for differentiation. Afterward, target cells were rinsed with 1X PBS, detached and used for experiments.

Kozlowska A.K., Topchyan P., Kaur K., Tseng H.C., Teruel A., Hiraga T, & Jewett A. (2017). Differentiation by NK cells is a prerequisite for effective targeting of cancer stem cells/poorly differentiated tumors by chemopreventive and chemotherapeutic drugs. Journal of Cancer, 8(4), 537-554.

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HepG2 Cells Differentiation and T-Cell Cytotoxicity

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HepG2 and HepG2.2.15 hepatoma cells were kept in full DMEM medium (DMEM, 10% FCS, 1% pen/strep, 1% sodium pyruvate, 1% NEAA; Life Technologies). For co-culture experiments 5x10⁴ target cells per well were seeded in collagen-coated (Serva, Heidelberg, Germany) 96-well flat bottom plates. When cells had reached confluence, differentiation medium was applied (Williams medium, 5% FCS, 1% Pen/Strep, 1% sodium pyruvate, 1% NEAA and 0.5% DMSO (Sigma-Aldrich). Target cells were used for experiments after 10 to 14 days of differentiation. The number of effector T cells/well was adjusted according to the transduction efficiency of each receptor to identical numbers of TCR-expressing cells. Viability of target cells was assessed using an XTT assay (Roche) and supernatants were subjected to an IFN-γ ELISA (BioLegend).

Wisskirchen K., Metzger K., Schreiber S., Asen T., Weigand L., Dargel C., Witter K., Kieback E., Sprinzl M.F., Uckert W., Schiemann M., Busch D.H., Krackhardt A.M, & Protzer U. (2017). Isolation and functional characterization of hepatitis B virus-specific T-cell receptors as new tools for experimental and clinical use. PLoS ONE, 12(8), e0182936.

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Cytokine-mediated T cell activation

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Unless otherwise specified,
reagents were used as received without further purification. Recombinant
human IL-2 (200-02, Peprotech), recombinant human IL-15 (570308, Biolegend),
recombinant mouse scIL-12 (130-096, Miltenyi Biotec), and recombinant
human IL-12 (CT050-HNAH, Sino Biological). Sulfo-Cyanine7 NHS ester
(Lumiprobe), DBCO-NHS ester (1160, Click Chemistry Tools), NHS-PEG-TCO
ester (A137-10, Click Chemistry Tools), poly(ethylene glycol)methyl
ether DBCO (20 kDa, A120-100, Click Chemistry Tools), and poly(ethylene
glycol)methyl ether azide (20 kDa, Nanocs). Polyacrylamide gels (Bio-Rad,
4–16 wt %). IFNγ ELISA (430104, Biolegend). CD19 antibody
(BD Biosciences Clone SJ25C1) and PD-L1 antibody (eBioscience Clone
MIH1).

Birnbaum L.A., Sullivan E.C., Do P., Uricoli B., Raikar S.S., Porter C.C., Henry C.J, & Dreaden E.C. (2023). Multicolor Light-Induced Immune Activation via Polymer Photocaged Cytokines. Biomacromolecules, 24(3), 1164-1172.

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Differentiation of Tumor Cells by Split Anergized NK Cells

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Freshly purified NK cells were treated with the combination of anti-CD16mAb (3 μg/mL) and IL-2 (1000 units/mL) for 18–24 h, then split anergized NK cell supernatants were harvested, and IFN-γ concentration in supernatants was determined using IFN-γ ELISA (Biolegend). Poorly differentiated tumor cells were differentiated using increasing amounts of split anergized NK cell supernatants added on a daily basis. Split anergized NK cell supernatants containing 1000 pg of IFN-γ were required to induce differentiation of 1 × 10⁶ OSCSCs while 35,000 pg of IFN-γ was needed to differentiate 1 × 10⁶ X01GB and X02GB over the course of 5–7 days. Differentiation status of each cell type was evaluated based on their susceptibility to NK cell-mediated cytotoxicity and surface expression of CD54, B7H1, MHC-I and CD44.

Kozlowska A.K., Tseng H.C., Kaur K., Topchyan P., Inagaki A., Bui V.T., Kasahara N., Cacalano N, & Jewett A. (2016). Resistance to cytotoxicity and sustained release of interleukin-6 and interleukin-8 in the presence of decreased interferon-γ after differentiation of glioblastoma by human natural killer cells. Cancer immunology, immunotherapy : CII, 65(9), 1085-1097.

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ELISA for IL-17 and IFN-γ Quantification

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Supernatants were collected from cell culture samples at the time of cell collection. Samples were retained at − 80 °C until ELISA was performed. IL-17 ELISA (BioLegend) was performed according to manufacturer’s instructions at sample dilutions of either 2× or 5×. IFN-γ ELISA (BioLegend) was performed according to manufacturer’s instructions at sample dilutions of either 50× or 100×.

Kiernan K., Alwarawrah Y., Nichols A.G., Danzaki K, & MacIver N.J. (2024). Insulin and IGF-1 have both overlapping and distinct effects on CD4+ T cell mitochondria, metabolism, and function. Scientific Reports, 14, 4331.

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